Bacterial antibiotic resistance levels in Danish farmland as a result of treatment with pig manure slurry

Resistance to tetracycline, macrolides and streptomycin was measured for a period of 8 months in soil bacteria obtained from farmland treated with pig manure slurry. This was done by spread plating bacteria on selective media (Luria Bertani (LB) medium supplemented with antibiotics). To account for seasonal variations in numbers of soil bacteria, ratios of resistant bacteria divided by total count on nonselective plates were calculated. Soil samples were collected from four different farms and from a control soil on a fifth farm. The control soil was not amended with animal manure. The occurrence of tetracycline-resistant bacteria was elevated after spread of pig manure slurry but declined throughout the sampling period to a level corresponding to the control soil. Higher load of pig manure slurry yielded higher occurrence of tetracycline resistance after spreading; however, the tetracycline resistance declined to normal occurrence defined by the tetracycline resistance occurrence in the control soil. Concentrations of tetracycline in soil and in pig manure slurry were measured using HPLC. No tetracycline exceeding the detection limit was found in soil samples. Manure slurry concentrations of tetracycline for three of the farms were 42, 81 and 698 mug/l, respectively. For streptomycin and macrolides, only minor variations in resistance levels were detected. Results obtained in this study thus indicate that tetracycline resistance levels in soil are temporarily influenced by the addition of pig manure slurry. The results indicate also that increased amount of pig manure slurry amendment may result in increased levels of tetracycline resistance in the soil. (C) 2002 Elsevier Science Ltd. All rights reserved.